A Convenient Nucleic Acid Test on the Basis of the Capillary Convective PCR for the On-Site Detection of Enterovirus 71

• Autores: Shiyin Zhang, Yanyan Lin, Jin Wang, Penglin Wang, Jieyu Chen, Miaoge Xue, Shuizhen He, Wenbin Zhou, Feihai Xu, Pingguo Liu, Pinghei Chen, Shengxiang Ge, Ningshao Xia
• Localización: The Journal of molecular diagnostics, ISSN 1525-1578, Vol. 16, Nº. 4, 2014, págs. 452-458
• Idioma: inglés
• DOI: 10.1016/j.jmoldx.2014.04.002
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• Resumen

  • The recent and continuing epidemic of enterovirus 71 in China has affected millions of children and resulted in thousands of deaths. Timely diagnosis and management is essential for disease control. Current enterovirus 71 molecular tests require resources that are unavailable for on-site testing. We have developed a simple-to-operate nucleic acid test, the convenient and integrated nucleic acid test, for local medical institutions. It uses a convective PCR for rapid amplification, a dipstick for visual detection of PCR products, and a simple commercial kit for nucleic acid extraction. By using a specially designed reagent and reaction tube containing a dipstick, the amplification and detection processes are well integrated and simplified. Moreover, cross contamination that may be caused by an open-tube detection system can be avoided. On the basis of the convenient and integrated nucleic acid test, an enterovirus 71 assay for on-site testing was developed. After evaluating known hand, foot, and mouth disease virus stocks of 17 strains of 11 different serotypes, this assay showed a favorable detection spectrum and no cross-reactivity. Its clinical performance was established by testing 141 clinical samples and comparing the results with a nested RT-PCR method. The assay showed a clinical sensitivity and specificity of 98.5% and 100%, respectively. Our results suggest that this convenient and integrated nucleic acid test enterovirus 71 assay may serve as an on-site diagnosis tool.